Obtaining and evaluating of enzymatic extracts from hairless canary seed (CDC Maria) as gluten-free bread-improving agents.

In this study, enzyme extracts were obtained from hairless canary seeds (CDC Maria) and used as gluten-free bread improvers. The enzyme extraction was done with a specific protein buffer solution and subsequent centrifugation. The supernatant was called crude enzyme extract, a fraction of this extract was refrigerated (CE) and another fraction was lyophilized (CEL). The lyoprotective effect of corn fiber (CEL + CF), maltodextrin (CEL + M), and inulin (CEL + I) was evaluated. Each enzyme extract was added to a gluten-free bread at 0.25%, 0.5%, 0.75% and 1% (w/w). The quality of the gluten-free bread was determined by external and internal characteristically, physical and sensory analysis: analysis of the lamella thickness, the shape factor of pores, the final volume, the aeration percentage, the texture profile analysis, the pore size distribution and shelf-life time. The results showed that the formulation with CEL at 0.5% (w/w) significantly improved the gluten-free bread quality properties, generating an increase of the final volume and aeration percentage, a reduction of the firmness, chewiness and sample aging, and a finer and more uniform crumb structure when compared to a control sample (P < 0.001). This study revealed the potential of a food-improving additive obtained from a natural origin with a high-level production in Argentina.

Trueperella pyogenes como causa de aborto em uma vaca - relato de caso / Trueperella pyogenes as an abortion cause in a cow - case report

Trueperella pyogenes é uma bactéria oportunista que causa mastite, metrite e abortos esporádicos em bovinos. Este trabalho relata um caso incomum de abort em uma vaca por Trueperella pyogenes. Um feto bovino, fêmea, mestiço Brahman, com oito meses de gestação, foi encaminhado para exame anatomopatológico e exames complementares. Na necropsia, evidenciou-se grande quantidade de líquido serossanguinolento e moderada quantidade de fibrina recobrindo a pleura visceral e o saco pericárdico. Os pulmões estavam difusamente avermelhados e consolidados, com áreas firmes esbranquiçadas ao corte de não mais de 1cm. No exame histopatológico, observou-se pneumonia necrossupurativa, pleurite fibrinopurulenta e placentite purulenta. No exame microbiológico, isolou-se T. pyogenes nas amostras de fígado, pulmões, conteúdo abomasal do feto e placenta. O feto foi negativo na PCR para Neospora caninum, Toxoplasma gondii e vírus da diarreia viral bovina (BVDV). Trueperella pyogenes geralmente causa broncopneumonia supurativa com formação de abscessos, porém, no presente feto abortado, observaram-se lesões macro e microscópicas comumente descritas em casos de aborto por Brucella abortus. Este estudo constata, então, a importância dessa bactéria como causa de aborto em bovinos, com lesões semelhantes à brucelose, destacando sua relevância dentro das causas de aborto em bovinos e o potencial zoonótico pouco explorado.(AU)

Trueperella pyogenes is an opportunistic bacterium associated with mastitis, metritis and occasional abortion in bovines. Here we report an uncommon case of abortion by T. pyogenes in a cow. An aborted female Brahman bovine fetus, at 8 months of gestational age was submitted for anatomopathological examination and complementary diagnostic tests. Macroscopic findings at necropsy included large amounts of free serum-blood fluid and moderate fibrin deposition covering both the visceral pleura and pericardial sack. The lungs were diffusely reddened and markedly consolidated, showing widespread smaller than 1cm, hard, white nodules. Necrosuppurative pneumonia, fibrinopurulent pleuritis, and purulent placentitis were the main histopathologic alterations observed. Trueperella pyogenes was isolated from liver, lungs, abomasa contents and placental samples. All tissue samples were PCR-negative for Neospora caninum, Toxoplasma gondii and bovine viral diarrhea virus (BVDV). Although T. pyogenes is often involved in suppurative bronchopneumonia and abscesses formation, macro and microscopic lesions in the present report were compatible with those commonly attributed to Brucella abortus fetal infections. Trueperella pyogenes is an important bovine pathogen with a neglected zoonotic potential being responsible for infections that can mimic other diseases' typical presentations.(AU)

Feline Leukaemia Virus Associated with Leukaemia in Cats in Santa Catarina, Brazil.

Leukaemia is a haemopoietic neoplasm originating from myeloid or lymphoid precursors in the bone marrow and may be either acute or chronic. These tumours are rare, but occur more frequently in cats because of an association with the feline leukaemia virus (FeLV) and feline immunodeficiency virus (FIV). To the best of our knowledge, no studies conducted in Brazil to date have analysed the association between leukaemia and FeLV and FIV infection in cats. The aim of this study was to perform a histopathological analysis of feline leukaemia and evaluate the association between leukaemia and FeLV and FIV infection in cats. The study evaluated 37 cats with leukaemia diagnosed between 2009 and 2017. The animals underwent necropsy examination, histopathology and immunohistochemistry with anti-FeLV gp70 and anti-FIV p24 gag antibodies. Of the evaluated animals, 54% (20/37) were males and 43.2% (16/37) were females. With respect to the life stage of the animals, 24.3% (9/37) were junior, 32.4% (12/37) were prime, 18.9% (7/37) were mature and 10.8% (4/37) were senior, and five animals were of unknown age. Myeloid leukaemia occurred in 56.8% (21/37) of the cases and lymphocytic leukaemia occurred in 43.2% (16/37) of the cases. Acute leukaemia (73%, 27/37) was more common than chronic leukaemia (27%, 10/37). The positivity for FeLV (78.4%, 29/37) and FIV (16.2%, 6/37) indicated a high association between FeLV infection and tumour development in the study region.

Feline Lymphoma and a High Correlation with Feline Leukaemia Virus Infection in Brazil.

Lymphoma is the most important haemopoietic tumour in cats and has been associated with feline leukaemia virus (FeLV) infection. In Brazil, no studies have established a correlation between FeLV infection and lymphoma. The aim of this study was to characterize lymphomas arising in cats in Brazil anatomically and microscopically, and to correlate these data with FeLV infection as determined by immunohistochemistry for the FeLV gp70 antigen. Fifty-three cats with lymphoma were evaluated. The mean age of junior, prime, mature, senior and geriatric cats was 1.65 years, 4.34 years, 8 years, 12.14 years and 15.5 years, respectively. The anatomical types of lymphoma were multicentric (43.4%, 23/53), mediastinal (33.96%, 18/53), renal (11.32%, 6/53), hepatic (5.66%, 3/53), nodal (3.77%, 2/53) and alimentary (1.89 %, 1/53). The histological types were small noncleaved-cell (33.96%, 18/53), mixed diffuse (22.64%, 12/53), immunoblastic (15.11%, 8/53), lymphoblastic (11, 32%, 6/53), small lymphocytic (9.43%, 5/53), small cleaved-cell (3.77%, 2/53) and large cell lymphomas (3.77%, 2/3). Immunopositivity for FeLV was observed in 56.6% (30/53) of the samples. FeLV positivity was equally distributed between the genders, but predominated in junior and prime cats. The degree of association between lymphoma and FeLV infection in Brazil was higher than that found in other countries, demonstrating the need to prevent and control the factors associated with infection.

Interference of age and supplementation of direct-fed microbial and essential oil in the activity of digestive enzymes and expression of genes related to transport and digestion of carbohydrates and proteins in the small intestine of broilers.

The objectives of this study were to describe alterations that age and dietary inclusion of direct-fed microbial (DFM) Bacillus subtilis (BS) and a specific essential oil (EO) blend (carvacrol, cinnamaldehyde, cineol, and pepper extract) causes in the activity of digestive enzymes (maltase: MALT; aminopeptidase-N: APN; intestinal alkaline phosphate: IAP) and expression patterns of genes related to transport (oligopeptide transporter gene: SLC15A1; Na+-dependent glucose and galactose transporter gene: SLC5A1; Na+-independent glucose, galactose, and fructose transporter gene: SLC2A2; ATPase, Na+/K+ transporting gene: ATP1A1) and digestion (aminopeptidase-N gene: ANPEP; maltase-glucoamylase gene: MGAM; Sucrase-isomaltase gene: SI) of carbohydrates and proteins in the small intestine of broilers. Also, the objective was to analyze if growth performance of broilers is affected by supplementation (BS and EO blend). Day-old male broiler chicks (n = 1,320) were assigned to 5 treatments. Diets included a basal diet (BD) as a negative control (CON); experimental diets were BD + BS; BD + BS + EO; BD + EO; BD + antibiotic growth promoter (AGP) avilamycin was the positive control. Performance was evaluated between 1 to 42 d. Transcript abundance of transport-related genes and digestion-related genes were assayed by RT-qPCR and determined at d 7, 21, and 42. MALT-, APN-, and IAP-specific activities were determined at d 7, 21, and 42. Broilers fed BS had greater SLC15A1 mRNA abundance compared to CON, while EO and AGP were related to higher activities of IAP and APN. Analysis over time revealed higher abundance of MGAM, SLC2A2, SLC15A1, SLC5A1 and SI mRNA at d 42 when compared to d 7. Activity of IAP decreased after d 7 and activity of MALT increased with age. The current study suggests that age had effect over carbohydrate and protein transport and carbohydrate digestion. The supplementation of BS DFM hade evident effect over protein transport and that the use of EO in the diet enhanced the activities of carbohydrate and protein digestion, reflecting improvement in digestive and transport physiology of birds. Changes performed by BS DFM and EO did not favor performance.

Risk assessment of maize damage by wireworms (Coleoptera: Elateridae) as the first step in implementing IPM and in reducing the environmental impact of soil insecticides.

A survey of maize fields was conducted in northeast Italy from 1986 to 2014, resulting in a dataset of 1296 records including information on wireworm damage to maize, plant-attacking species, agronomic characteristics, landscape and climate. Three wireworm species, Agriotes brevis Candeze, A. sordidus Illiger and A. ustulatus Schäller, were identified as the dominant pest species in maize fields. Over the 29-year period surveyed, no yield reduction was observed when wireworm plant damage was below 15 % of the stand. A preliminary univariate analysis of risk assessment was applied to identify the main factors influencing the occurrence of damage. A multifactorial model was then applied by using the significant factors identified. This model allowed the research to highlight the strongest factors and to analyse how the main factors together influenced damage risk. The strongest factors were: A. brevis as prevalent damaging species, soil organic matter content >5 %, rotation including meadows and/or double crops, A. sordidus as prevalent damaging species, and surrounding landscape mainly meadows, uncultivated grass and double crops. The multifactorial model also showed how the simultaneous occurrence of two or more of the aforementioned risk factors can conspicuously increase the risk of wireworm damage to maize crops, while the probability of damage to a field with no-risk factors is always low (<1 %). These results make it possible to draw risk maps to identify low-risk and high-risk areas, a first step in implementing bespoke IPM procedures in an attempt to reduce the impact of soil insecticides significantly.

Systemic insecticides (neonicotinoids and fipronil): trends, uses, mode of action and metabolites.

Since their discovery in the late 1980s, neonicotinoid pesticides have become the most widely used class of insecticides worldwide, with large-scale applications ranging from plant protection (crops, vegetables, fruits), veterinary products, and biocides to invertebrate pest control in fish farming. In this review, we address the phenyl-pyrazole fipronil together with neonicotinoids because of similarities in their toxicity, physicochemical profiles, and presence in the environment. Neonicotinoids and fipronil currently account for approximately one third of the world insecticide market; the annual world production of the archetype neonicotinoid, imidacloprid, was estimated to be ca. 20,000 tonnes active substance in 2010. There were several reasons for the initial success of neonicotinoids and fipronil: (1) there was no known pesticide resistance in target pests, mainly because of their recent development, (2) their physicochemical properties included many advantages over previous generations of insecticides (i.e., organophosphates, carbamates, pyrethroids, etc.), and (3) they shared an assumed reduced operator and consumer risk. Due to their systemic nature, they are taken up by the roots or leaves and translocated to all parts of the plant, which, in turn, makes them effectively toxic to herbivorous insects. The toxicity persists for a variable period of time-depending on the plant, its growth stage, and the amount of pesticide applied. A wide variety of applications are available, including the most common prophylactic non-Good Agricultural Practices (GAP) application by seed coating. As a result of their extensive use and physicochemical properties, these substances can be found in all environmental compartments including soil, water, and air. Neonicotinoids and fipronil operate by disrupting neural transmission in the central nervous system of invertebrates. Neonicotinoids mimic the action of neurotransmitters, while fipronil inhibits neuronal receptors. In doing so, they continuously stimulate neurons leading ultimately to death of target invertebrates. Like virtually all insecticides, they can also have lethal and sublethal impacts on non-target organisms, including insect predators and vertebrates. Furthermore, a range of synergistic effects with other stressors have been documented. Here, we review extensively their metabolic pathways, showing how they form both compound-specific and common metabolites which can themselves be toxic. These may result in prolonged toxicity. Considering their wide commercial expansion, mode of action, the systemic properties in plants, persistence and environmental fate, coupled with limited information about the toxicity profiles of these compounds and their metabolites, neonicotinoids and fipronil may entail significant risks to the environment. A global evaluation of the potential collateral effects of their use is therefore timely. The present paper and subsequent chapters in this review of the global literature explore these risks and show a growing body of evidence that persistent, low concentrations of these insecticides pose serious risks of undesirable environmental impacts.

Analysis of the intestinal bacterial microbiota in maize- or sorghum-fed broiler chickens using real-time PCR.

1. An experiment was conducted to study the effect of two different diets on zootechnical performance and the major bacterial groups in association with the host mucosa and dispersed in the lumen contents of the small intestine of broiler chickens. 2. The two experimental diets were maize or sorghum-based. In addition to the total bacteria, bacterial groups belonging to the Enterobacteriaceae (Enterococcus and Lactobacillus) were quantified by real-time PCR. 3. There were no differences in body weight gain and feed intake, but feed conversion ratio increased for sorghum-fed broilers at 21 and 42 d of age. 4. The Enterococcus group decreased in all gut segments from 7 to 42 d, while the Lactobacillus group increased in both ecosystems. In the ileal mucosa, the enterobacterial counts decreased from 7 to 42 d in the maize-based diet, but remained stable in the sorghum-based diet. 5. The results shed light on the spatial and temporal distribution of bacterial groups that play important physiological roles in the small intestine of chickens. Specifically, the increased Enterobacteria population in the ileum is consistent with the relatively poor feed conversion in sorghum-fed broilers.

Predictive accuracy of triage nurses evaluation in risk stratification of syncope in the emergency department.

BACKGROUND: Syncope is a common clinical problem that accounts for 1-3% of all emergency department (ED) visits. Its prognosis is extremely variable with a 1-year mortality that may reach 30%. There are no available data about the accuracy of nursing triage in identifying high-risk syncope. The aim of our study was to evaluate the predictive accuracy of nursing triage in identifying high-risk syncope. METHODS: We conducted a retrospective study on 678 consecutive patients who presented with syncope at four EDs. For each patient, nursing triage, comorbidities, clinical features and adverse events that occurred both in the ED and at 10-day follow-up were assessed. Adverse events included death, readmission to ED, need for major therapeutic procedures, cardiopulmonary resuscitation, intensive care unit admittance, acute antiarrhythmic therapy and major causes of syncope identified during the ED evaluation. Predictive accuracy of nursing triage was evaluated. RESULTS: We observed a total of 55 (8.1%) adverse events. Eight of them (9.4%) occurred among the 85 patients who were identified as high priority by nursing triage. Sensitivity (Sn) and specificity (Sp) of urgent nursing triage in identifying adverse outcomes in the ED (19 patients) were 21% (95% CI 3% to 39%) and 88% (95% CI 85% to 90%), while the positive likelihood ratio (LR+) and negative likelihood ratio (LR-) were 1.7 and 0.9, respectively. Sn and Sp for 10-day adverse events were 15% (95% CI 5% to 24%) and 88% (95% CI 85% to 90%), respectively, with a LR+ of 1.18 and a LR- of 0.98. CONCLUSIONS: Nursing triage was characterised by a low predictive accuracy in identifying high-risk individuals.

PCR-based species identification of Agriotes larvae.

Click beetle larvae within the genus Agriotes (Coleoptera: Elateridae), commonly known as wireworms, are abundant ground-dwelling herbivores which can inflict considerable damage to field crops. In Central Europe up to 20 species, which differ in their distribution, ecology and pest status, occur in arable land. However, the identification of these larvae based on morphological characters is difficult or impossible. This hampers progress towards controlling these pests. Here, we present a polymerase chain reaction (PCR)-based approach to identify, for the first time, 17 Agriotes species typically found in Central Europe. Diagnostic sequence information was generated and submitted to GenBank, allowing the identification of these species via DNA barcoding. Moreover, multiplex PCR assays were developed to identify the nine most abundant species rapidly within a single-step reaction: Agriotes brevis, A. litigiosus, A. obscurus, A. rufipalpis, A. sordidus, A. sputator, A. ustulatus, A. lineatus and A. proximus. The latter two species remain molecularly indistinguishable, questioning their species status. The multiplex PCR assays proved to be highly specific against non-agrioted elaterid beetles and other non-target soil invertebrates. By testing the molecular identification system with over 900 field-collected larvae, our protocol proved to be a reliable, cheap and quick method to routinely identify Central European Agriotes species.

Identification and characterization of polymorphisms within the 5' flanking region, first exon and part of first intron of bovine GH gene.

The aim of the present study was to identify and characterize polymorphisms within the 5' flanking region, first exon and part of first intron of the bovine growth hormone gene among different beef cattle breeds: Nelore (n = 25), Simmental (n = 39), Simbrasil (n = 24), Simmental x Nelore (n = 30), Canchim x Nelore (n = 30) and Angus x Nelore (n = 30). Two DNA fragments (GH1, 464 bp and GH2, 453 bp) were amplified by polymerase chain reaction and then used for polymorphism identification by SSCP. Within the GH1 fragment, five polymorphisms were identified, corresponding to three different alleles: GH1.1, GH1.2 and GH1.3 (GenBank: AY662648, AY662649 and AY662650, respectively). These allele sequences were aligned and compared with bovine GH gene nucleotide sequence (GenBank: M57764 and AF118837), resulting in the identification of five insertion/deletions (INDELs) and five single nucleotide polymorphisms (SNPs). In the GH2 fragment two alleles were identified, GH2.1 and GH2.2 (GenBank: AY662651 and AY662652, respectively). The allele sequences were compared with GenBank sequences (M57764, AF007750 and AH009106) and three INDELs and four SNPs were identified. In conclusion, we were able to identify six new polymorphisms of the bovine GH gene (one INDEL and five SNPs), which can be used as molecular markers in genetic studies.

Comparative genomics of two Leptospira interrogans serovars reveals novel insights into physiology and pathogenesis.

Leptospira species colonize a significant proportion of rodent populations worldwide and produce life-threatening infections in accidental hosts, including humans. Complete genome sequencing of Leptospira interrogans serovar Copenhageni and comparative analysis with the available Leptospira interrogans serovar Lai genome reveal that despite overall genetic similarity there are significant structural differences, including a large chromosomal inversion and extensive variation in the number and distribution of insertion sequence elements. Genome sequence analysis elucidates many of the novel aspects of leptospiral physiology relating to energy metabolism, oxygen tolerance, two-component signal transduction systems, and mechanisms of pathogenesis. A broad array of transcriptional regulation proteins and two new families of afimbrial adhesins which contribute to host tissue colonization in the early steps of infection were identified. Differences in genes involved in the biosynthesis of lipopolysaccharide O side chains between the Copenhageni and Lai serovars were identified, offering an important starting point for the elucidation of the organism's complex polysaccharide surface antigens. Differences in adhesins and in lipopolysaccharide might be associated with the adaptation of serovars Copenhageni and Lai to different animal hosts. Hundreds of genes encoding surface-exposed lipoproteins and transmembrane outer membrane proteins were identified as candidates for development of vaccines for the prevention of leptospirosis.

Comparative analyses of the complete genome sequences of Pierce's disease and citrus variegated chlorosis strains of Xylella fastidiosa.

Xylella fastidiosa is a xylem-dwelling, insect-transmitted, gamma-proteobacterium that causes diseases in many plants, including grapevine, citrus, periwinkle, almond, oleander, and coffee. X. fastidiosa has an unusually broad host range, has an extensive geographical distribution throughout the American continent, and induces diverse disease phenotypes. Previous molecular analyses indicated three distinct groups of X. fastidiosa isolates that were expected to be genetically divergent. Here we report the genome sequence of X. fastidiosa (Temecula strain), isolated from a naturally infected grapevine with Pierce's disease (PD) in a wine-grape-growing region of California. Comparative analyses with a previously sequenced X. fastidiosa strain responsible for citrus variegated chlorosis (CVC) revealed that 98% of the PD X. fastidiosa Temecula genes are shared with the CVC X. fastidiosa strain 9a5c genes. Furthermore, the average amino acid identity of the open reading frames in the strains is 95.7%. Genomic differences are limited to phage-associated chromosomal rearrangements and deletions that also account for the strain-specific genes present in each genome. Genomic islands, one in each genome, were identified, and their presence in other X. fastidiosa strains was analyzed. We conclude that these two organisms have identical metabolic functions and are likely to use a common set of genes in plant colonization and pathogenesis, permitting convergence of functional genomic strategies.

Growth hormone mRNA expression in the pituitary of Bos indicus and Bos taurus x Bos indicus crossbred young bulls treated with recombinant bovine somatotropin

The effects of breed and of recombinant bovine somatotropin (rbST) treatment on growth hormone gene expression were studied in young bulls. The experiment was completely randomized in a [2 x 2]-factorial arrangement, using two levels of rbST (0 or 250 mg/animal/14 days), and two breed groups (Nelore and Simmental x Nelore crossbred). A cDNA encoding Bos indicus growth hormone was cloned and sequenced for use as a probe in Northern and dot blot analyses. Compared to the Bos taurus structural gene, the Bos indicus cDNA was found to begin 21 bases downstream from the transcription initiation site and had only two discrepancies (C to T at position 144-His and T to C at position 354-Phe), without changes in the polypeptide sequence. However, two amino acid substitutions were found for Bubalus spp., which belong to the same tribe. The rbST treatment did not change any of the characteristics evaluated (body and pituitary gland weights, growth hormone mRNA expression level). Crossbred animals had significantly higher body weight and heavier pituitaries than Nelore cattle. Pituitary weight was proportional to body weight in both breed groups. Growth hormone mRNA expression in the pituitary was similar (P>0.075) for both breed and hormonal treatment groups, but was 31.9 higher in the pure Nelore group, suggesting that growth hormone gene transcription regulation differs among these breeds

Comparison of the genomes of two Xanthomonas pathogens with differing host specificities.

The genus Xanthomonas is a diverse and economically important group of bacterial phytopathogens, belonging to the gamma-subdivision of the Proteobacteria. Xanthomonas axonopodis pv. citri (Xac) causes citrus canker, which affects most commercial citrus cultivars, resulting in significant losses worldwide. Symptoms include canker lesions, leading to abscission of fruit and leaves and general tree decline. Xanthomonas campestris pv. campestris (Xcc) causes black rot, which affects crucifers such as Brassica and Arabidopsis. Symptoms include marginal leaf chlorosis and darkening of vascular tissue, accompanied by extensive wilting and necrosis. Xanthomonas campestris pv. campestris is grown commercially to produce the exopolysaccharide xanthan gum, which is used as a viscosifying and stabilizing agent in many industries. Here we report and compare the complete genome sequences of Xac and Xcc. Their distinct disease phenotypes and host ranges belie a high degree of similarity at the genomic level. More than 80% of genes are shared, and gene order is conserved along most of their respective chromosomes. We identified several groups of strain-specific genes, and on the basis of these groups we propose mechanisms that may explain the differing host specificities and pathogenic processes.

Growth hormone mRNA expression in the pituitary of Bos indicus and Bos taurus x Bos indicus crossbred young bulls treated with recombinant bovine somatotropin.

The effects of breed and of recombinant bovine somatotropin (rbST) treatment on growth hormone gene expression were studied in young bulls. The experiment was completely randomized in a [2 x 2]-factorial arrangement, using two levels of rbST (0 or 250 mg/animal/14 days), and two breed groups (Nelore and Simmental x Nelore crossbred). A cDNA encoding Bos indicus growth hormone was cloned and sequenced for use as a probe in Northern and dot blot analyses. Compared to the Bos taurus structural gene, the Bos indicus cDNA was found to begin 21 bases downstream from the transcription initiation site and had only two discrepancies (C to T at position 144-His and T to C at position 354-Phe), without changes in the polypeptide sequence. However, two amino acid substitutions were found for Bubalus spp., which belong to the same tribe. The rbST treatment did not change any of the characteristics evaluated (body and pituitary gland weights, growth hormone mRNA expression level). Crossbred animals had significantly higher body weight and heavier pituitaries than Nelore cattle. Pituitary weight was proportional to body weight in both breed groups. Growth hormone mRNA expression in the pituitary was similar (P>0.075) for both breed and hormonal treatment groups, but was 31.9% higher in the pure Nelore group, suggesting that growth hormone gene transcription regulation differs among these breeds.

The genome sequence of the plant pathogen Xylella fastidiosa. The Xylella fastidiosa Consortium of the Organization for Nucleotide Sequencing and Analysis.

Xylella fastidiosa is a fastidious, xylem-limited bacterium that causes a range of economically important plant diseases. Here we report the complete genome sequence of X. fastidiosa clone 9a5c, which causes citrus variegated chlorosis--a serious disease of orange trees. The genome comprises a 52.7% GC-rich 2,679,305-base-pair (bp) circular chromosome and two plasmids of 51,158 bp and 1,285 bp. We can assign putative functions to 47% of the 2,904 predicted coding regions. Efficient metabolic functions are predicted, with sugars as the principal energy and carbon source, supporting existence in the nutrient-poor xylem sap. The mechanisms associated with pathogenicity and virulence involve toxins, antibiotics and ion sequestration systems, as well as bacterium-bacterium and bacterium-host interactions mediated by a range of proteins. Orthologues of some of these proteins have only been identified in animal and human pathogens; their presence in X. fastidiosa indicates that the molecular basis for bacterial pathogenicity is both conserved and independent of host. At least 83 genes are bacteriophage-derived and include virulence-associated genes from other bacteria, providing direct evidence of phage-mediated horizontal gene transfer.

Concomitant radiotherapy with mitomycin C and bleomycin compared with radiotherapy alone in inoperable head and neck cancer: final report.

PURPOSE: To compare the efficacy of concomitant irradiation with mitomycin C and bleomycin in patients with inoperable head and neck carcinoma with radiotherapy alone. METHODS AND MATERIALS: Between March 1991 and December 1993, 64 patients with inoperable head and neck carcinoma (41 with oropharyngeal site) were randomized to radiotherapy alone (group A) or radiotherapy combined with simultaneous application of mitomycin C and bleomycin (group B). In both groups patients were irradiated five times weekly with 2 Gy to a total dose of 66-70 Gy. The planned concomitant treatment in group B was: bleomycin 5 units twice a week i.m., total dose 70 units, mitomycin C 15 mg/m2 i.v. after delivery of 10 Gy, and 10 mg/m2 i.v. on the last day of radiotherapy. To enhance the effect of these two drugs, patients received also nicotinamide, chlorpromazine, and dicoumarol. Because significantly better results were achieved in arm B for patients with inoperable oropharyngeal carcinoma, the study was closed and such patients were after December 1993 routinely treated with the combined therapy (as in arm B). Until October 1996, we treated and followed up 48 such consecutive patients. RESULTS: Median follow-up of our study patients is 42 months. Complete remission (CR) rate in group A was 31% and in group B 59% (p = 0.04); disease-free survival (DFS) in group A was 8% and in group B 37% (P = 0.01); and overall survival (OS) was 7% in group A and 26% in group B (p = 0.08). CR rate for patients with oropharyngeal carcinoma was 29% in group A (N = 21) and 75% in group B (N = 20) (p = 0.007); DFS in group A was 10% and in group B 48% (p = 0.001); and the OS was 10% in group A and 38% in group B (p = 0.019). In patients with inoperable oropharyngeal carcinoma treated after December 1993, complete remission was achieved in 32/48 (67%, 95% CI: 52%-80%). DFS at the median follow-up of 14 months was 60% (95% CI 43-77%) and OS 58% (95% CI 42-74%). CONCLUSION: From the results of our study it seems that the concomitant treatment significantly improves CR rate, DFS, and OS in patients with inoperable oropharyngeal carcinoma in comparison with radiotherapy alone.

Lipid modifications of microsomes isolated from villus and crypt zones of bovine intestinal mucosa: relationship with fatty acid binding protein.

The present studies were conducted to examine the fatty acid composition of microsomal lipids of bovine small intestine. Microsomes and cytosol were isolated from mucosal scrapings enriched with villus and crypts cells, and the following studies were conducted to: 1) analyse fatty acids from microsomal lipids; 2) incorporate 1-14C oleic acid to microsomal lipids; and 3) bind 1-14C oleic acid to cytosolic proteins of villus and crypt zone. The results of these studies demonstrated that: (1) the major unsaturated fatty acids of microsomes were oleic (C18:1 n-9), linoleic (C18:2 n-6) and arachidonic acids (C20:4 n-6), which increased from crypt to villus tip of the bovine intestinal mucosa; (2) gel filtration chromatography indicated that the low-molecular weight cytosolic proteins obtained from superficial mucosal scrapings contained the greatest oleic acid binding activity; (3) the incorporation of 1-14C oleic acid to microsomes was higher in phospholipids, triglycerides and cholesterol esters from villus than in crypts zones; (4) the protein content of cytosol and microsomes was longer in villus zones than in crypts zones; (5) the peroxidizability index showed the highest value in villus microsomes.